descriptionPublicationkeyboard_double_arrow_right Article 06 Aug 2009 English Publisher:Springer Science and Business Media LLCJournal:Nature Protocols, volume 4, pages 1,245-1,251 (issn: 1754-2189, eissn: 1750-2799,

Authors: F. Peter Guengerich; Martha V. Martin; Christal D. Sohl; Qian Cheng;

doi: 10.1038/nprot.2009.121

pmid: 19661994

pmc: PMC3843963

Measurement of cytochrome P450 and NADPH–cytochrome P450 reductase

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Abstract

Cytochrome P450 (P450) enzymes are important in the metabolism of steroids, vitamins, carcinogens, drugs and other compounds. Two of the commonly used assays in this field are the measurements of total P450 and NADPH-P450 reductase in biological preparations. A detailed protocol is presented for the measurement of P450 by its spectral properties, along with a protocol for measuring NADPH-P450 reductase by its NADPH-cytochrome c reduction activity. Each assay can be completed in 5-10 min. Detailed explanations for the rationale of particular sequences in the protocols are provided, along with potential confounding problems.

Related Organizations

Vanderbilt University
United States

Keywords

Cytochrome P-450 Enzyme System, Spectrophotometry, Microsomes, Liver, Animals, Humans, NADPH-Ferrihemoprotein Reductase, Rats

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